Structural insights into the substrate specificity of a glycoside hydrolase family 5 lichenase from Caldicellulosiruptor sp F32

doi:10.1042/BCJ20170328

CORC > 青岛生物能源与过程研究所 > 中国科学院青岛生物能源与过程研究所 > 微生物资源团队

	Structural insights into the substrate specificity of a glycoside hydrolase family 5 lichenase from Caldicellulosiruptor sp F32
	Meng, Dong-Dong 1,5; Liu, Xi 2; Dong, Sheng 1,3,4; Wang, Ye-Fei 1; Ma, Xiao-Qing 1; Zhou, Haixia 2; Wang, Xinquan 2; Yao, Li-Shan 1; Feng, Yingang 1,3,4; Li, Fu-Li 1
刊名	BIOCHEMICAL JOURNAL
	2017-10-15
卷号	474 页码:3373-3389
DOI	10.1042/BCJ20170328
文献子类	Article
英文摘要	Glycoside hydrolase (GH) family 5 is one of the largest GH families with various GH activities including lichenase, but the structural basis of the GH5 lichenase activity is still unknown. A novel thermostable lichenase F32EG5 belonging to GH5 was identified from an extremely thermophilic bacterium Caldicellulosiruptor sp. F32. F32EG5 is a bi-functional cellulose and a lichenan-degrading enzyme, and exhibited a high activity on beta-1,3-1,4-glucan but side activity on cellulose. Thin-layer chromatography and NMR analyses indicated that F32EG5 cleaved the beta-1,4 linkage or the beta-1,3 linkage while a 4-O-substitued glucose residue linked to a glucose residue through a beta-1,3 linkage, which is completely different from extensively studied GH16 lichenase that catalyses strict endo-hydrolysis of the beta-1,4-glycosidic linkage adjacent to a 3-O-substitued glucose residue in the mixed-linked beta-glucans. The crystal structure of F32EG5 was determined to 2.8 angstrom resolution, and the crystal structure of the complex of F32EG5 E193Q mutant and cellotetraose was determined to 1.7 angstrom resolution, which revealed that the exit subsites of substrate-binding sites contribute to both thermostability and substrate specificity of F32EG5. The sugar chain showed a sharp bend in the complex structure, suggesting that a substrate cleft fitting to the bent sugar chains in lichenan is a common feature of GH5 lichenases. The mechanism of thermostability and substrate selectivity of F32EG5 was further demonstrated by molecular dynamics simulation and site-directed mutagenesis. These results provide biochemical and structural insights into thermostability and substrate selectivity of GH5 lichenases, which have potential in industrial processes.
WOS关键词	CRYSTAL-STRUCTURE ; 1,3-1,4-BETA-D-GLUCAN 4-GLUCANOHYDROLASE ; CLOSTRIDIUM-CELLULOVORANS ; FIBROBACTER-SUCCINOGENES ; ESCHERICHIA-COLI ; BETA-GLUCAN ; PROTEIN ; ENDOGLUCANASE ; XYLANASES ; COMPLEX
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
WOS记录号	WOS:000413116500001
资助机构	National Basic Research Program of China(2011CB707404) ; National Natural Science Foundation of China(31270784 ; Shandong Province Natural Science Funds for Distinguished Young Scholar(JQ201507) ; Key Scientific and Technological Project of Shandong Province(2015ZDXX0403A01) ; 31670735)
内容类型	期刊论文
源URL	[http://ir.qibebt.ac.cn/handle/337004/9562]
专题	青岛生物能源与过程研究所_微生物资源团队
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Qingdao 266101, Peoples R China 2.Tsinghua Univ, Sch Life Sci, Struct Biol Ctr, Minist Educ,Key Lab Prot Sci, Beijing 100084, Peoples R China 3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Energy Genet, Qingdao 266101, Peoples R China 4.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao Engn Lab Single Cell Oil, Qingdao 266101, Peoples R China 5.Univ Chinese Acad Sci, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Meng, Dong-Dong,Liu, Xi,Dong, Sheng,et al. Structural insights into the substrate specificity of a glycoside hydrolase family 5 lichenase from Caldicellulosiruptor sp F32[J]. BIOCHEMICAL JOURNAL,2017,474:3373-3389.
APA	Meng, Dong-Dong.,Liu, Xi.,Dong, Sheng.,Wang, Ye-Fei.,Ma, Xiao-Qing.,...&Li, Fu-Li.(2017).Structural insights into the substrate specificity of a glycoside hydrolase family 5 lichenase from Caldicellulosiruptor sp F32.BIOCHEMICAL JOURNAL,474,3373-3389.
MLA	Meng, Dong-Dong,et al."Structural insights into the substrate specificity of a glycoside hydrolase family 5 lichenase from Caldicellulosiruptor sp F32".BIOCHEMICAL JOURNAL 474(2017):3373-3389.